JP3411112B2 - Particle image analyzer - Google Patents
Particle image analyzerInfo
- Publication number
- JP3411112B2 JP3411112B2 JP27145394A JP27145394A JP3411112B2 JP 3411112 B2 JP3411112 B2 JP 3411112B2 JP 27145394 A JP27145394 A JP 27145394A JP 27145394 A JP27145394 A JP 27145394A JP 3411112 B2 JP3411112 B2 JP 3411112B2
- Authority
- JP
- Japan
- Prior art keywords
- particle
- image
- particles
- particle size
- circularity
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1468—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle
- G01N15/147—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle the analysis being performed on a sample stream
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/02—Investigating particle size or size distribution
- G01N15/0205—Investigating particle size or size distribution by optical means
- G01N15/0227—Investigating particle size or size distribution by optical means using imaging; using holography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/02—Investigating particle size or size distribution
- G01N2015/0294—Particle shape
Landscapes
- Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Dispersion Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Image Processing (AREA)
- Length Measuring Devices By Optical Means (AREA)
- Image Analysis (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Description
【０００１】[0001]
【産業上の利用分野】この発明は液中の粒子を撮像し、
その粒子像を記憶、表示するとともに、粒子像を画像解
析することによって、粒子の大きさや形状に関する情報
を求める粒子画像分析装置に関する。BACKGROUND OF THE INVENTION The present invention is for imaging particles in a liquid,
The present invention relates to a particle image analyzer that stores and displays the particle image and analyzes the image of the particle to obtain information regarding the size and shape of the particle.
【０００２】[0002]
【従来の技術】ファインセラミックス粒子、顔料、化粧
品用パウダー等の粉体の品質を管理する上で、粒子の粒
径を測定、管理することは非常に重要である。その測定
装置として、古くから液相沈降法、電気的検知帯法（ク
ールター法）による測定装置があり、最近ではレーザ回
析散乱法による測定装置が広く使用されている。2. Description of the Related Art In controlling the quality of fine ceramic particles, pigments, cosmetic powders and the like, it is very important to measure and control the particle size of the particles. As a measuring device therefor, a liquid phase sedimentation method and an electric detection zone method (Coulter method) have been used for a long time, and recently, a laser diffraction scattering method has been widely used.
【０００３】[0003]
【発明が解決しようとする課題】しかしながら、上記の
いづれの方式による測定装置においても、その測定精度
（正確度）は、いまだに満足できるものではない。特
に、対象とする粒子が偏平であったり細長い形をしてい
る場合には、測定方法の違いによって、求められる粒径
は大きく異なることがある。また、一般的に微小な粒子
は測定中に凝集しやすく、その場合にも正確な粒度分布
を求めることができない。また、粒子の球形度（円形
度）や凝集度合い等に関する情報を、上記従来の粒度分
布測定装置で得ることは困難である。However, the measurement accuracy (accuracy) of any of the above measuring devices is still unsatisfactory. In particular, when the particles of interest are flat or have an elongated shape, the required particle size may differ greatly depending on the measurement method. In addition, fine particles generally tend to agglomerate during measurement, and even in that case, an accurate particle size distribution cannot be obtained. In addition, it is difficult to obtain information about the sphericity (circularity) and the degree of aggregation of particles with the above-described conventional particle size distribution measuring device.
【０００４】懸濁液中の粒子の内、大きい粒子のほうが
小さい粒子に比べて速く沈降するので、粒子濃度は時間
的、空間的に変化する。この変化を光の透過量で検知し
て粒度分布を求める方法が、沈降法として代表的な液相
沈降光透過法である。この沈降法では、同じ体積と密度
の粒子でも、その粒子の形状が異なると沈降速度は異な
る。また、粒子どうしが凝集していると、その凝集粒子
は速く沈降する。Among the particles in the suspension, the larger particles settle faster than the smaller particles, so that the particle concentration changes temporally and spatially. A liquid phase sedimentation light transmission method, which is a typical sedimentation method, is a method of obtaining the particle size distribution by detecting this change by the amount of light transmission. In this sedimentation method, even particles having the same volume and density have different sedimentation rates if the shapes of the particles are different. Also, when the particles are agglomerated, the agglomerated particles settle quickly.
【０００５】電気的検知帯法による装置は、電解液に浮
遊させた粒子が小さな穴を通過する時の電気抵抗の変化
を検出するものであり、１個１個の粒子の体積相当径が
形状にほとんど影響されずに測定できる。逆に言えば、
電気的検知帯法では、粒子の形状に関する情報を得るこ
とは困難である。また、粒子の大きさと比較して電気的
検知領域がかなり広いので、粒子どうしが近接あるいは
凝集していると、正確な粒度分布を求めることができな
い。The device based on the electrical detection zone method detects changes in electric resistance when particles suspended in an electrolytic solution pass through a small hole, and each particle has a volume equivalent diameter. Can be measured with almost no effect. Conversely,
It is difficult to obtain information on the shape of particles by the electrical detection zone method. Further, since the electrical detection region is considerably wider than the size of the particles, an accurate particle size distribution cannot be obtained if the particles are close to each other or are agglomerated.
【０００６】最近広く使用されているレーザ回析散乱法
の装置は、浮遊している粒子群にレーザ光を照射して得
られる回析光／散乱光強度の角度分布情報から、ミー散
乱理論に基づいて粒径分布を推定、算出するものであ
る。この装置では粒度が未知の試料や屈折率が同じ粒子
の混合試料でも、粒径が０．１μｍから数百μｍまでの
粒子に対し、１回の測定で再現性のある粒度分布が得ら
れるという利点がある。The laser diffraction / scattering method that has been widely used recently is based on the Mie scattering theory based on the angular distribution information of the diffracted light / scattered light intensity obtained by irradiating a floating particle group with laser light. Based on this, the particle size distribution is estimated and calculated. With this device, it is possible to obtain a reproducible particle size distribution in a single measurement for particles with a particle size of 0.1 μm to several hundreds of μm even for samples of unknown particle size or mixed samples of particles with the same refractive index. There are advantages.
【０００７】しかし、この方式の装置には次に挙げるよ
うな問題点がある。
１）粒子による散乱光強度は、形状、屈折率、表面状態
等の違いによる影響を大きく受け、正確な粒度分布を求
めるのは難しい。
２）測定する粒子の正確な屈折率を入力する必要がある
が、粒子の表面が酸化していたり、不純物が混ざってい
ることがあり、文献値を入力しても正しく粒度分布が求
められないことがある。However, this type of device has the following problems. 1) The intensity of scattered light by particles is greatly affected by differences in shape, refractive index, surface state, etc., and it is difficult to obtain an accurate particle size distribution. 2) It is necessary to enter the correct refractive index of the particles to be measured, but the particle surface may be oxidized or impurities may be mixed, and the particle size distribution cannot be obtained correctly even if the literature values are entered. Sometimes.
【０００８】３）粒子が球形で表面が滑らかであり凝集
していないという仮定のもとに、多数の粒子による回析
光／散乱光強度分布についての連立方程式を解いて粒度
分布を推定する。その仮定を満足しない粒子に対して
は、その連立方程式が満足に解けないことがあり、独自
の補正を行っている。
４）上記のような独自の補正のために、機種間の測定結
果に大きな差が生じることがある。
以上のように、従来の粒度分布測定装置では、粒子の形
状や凝集の影響を大きく受け、正確な粒度分布を求める
のは難しい。また、粒子の形状や凝集度合いに関する情
報を得ることも困難である。3) The particle size distribution is estimated by solving a simultaneous equation for the diffracted light / scattered light intensity distribution by a large number of particles under the assumption that the particles are spherical and the surface is smooth and does not aggregate. For the particles that do not satisfy the assumption, the simultaneous equations may not be solved satisfactorily, and the original correction is performed. 4) Due to the above-described unique correction, a large difference may occur in the measurement results between models. As described above, in the conventional particle size distribution measuring device, it is difficult to obtain an accurate particle size distribution because it is greatly affected by the shape and aggregation of particles. Further, it is difficult to obtain information on the shape of particles and the degree of aggregation.
【０００９】粒子の形状を測定する方法としては、顕微
鏡と画像処理装置を組み合わせる方法がある。しかし、
工業用の粉体は、粉砕して作られた粒子が多く、そのよ
うな粉体では、ひとつの試料でも各粒子の大きさが著し
く異なり、スライドグラス上の粒子の全てにピントを合
わすことはできない。すなわち、小さな粒子に対してピ
ントを合わすと大きな粒子に対してピントが合わなくな
る。大きな粒子に対してピントを合わすと小さな粒子に
対してピントが合わなくなる。従って、この顕微鏡方式
は、粒子の大きさが揃っている場合にしか利用できない
方法である。As a method of measuring the shape of particles, there is a method of combining a microscope and an image processing apparatus. But,
Many industrial powders are made by crushing.In such powders, the size of each particle is significantly different even in one sample, and it is impossible to focus all the particles on the slide glass. Can not. That is, if a small particle is in focus, a large particle is out of focus. If you focus on a large particle, you will lose focus on a small particle. Therefore, this microscope method is a method that can be used only when the particle sizes are uniform.
【００１０】また、この顕微鏡方式で何千個もの粒子像
を解析しようとすると、撮像する視野を変更するために
スライドグラスを少しずつ移動させて何百枚も画像を取
り込んで解析する必要があり、手間と時間がかかる。こ
のような理由で、工業用粉体に対しては、粒子像から粒
子の大きさや形状を測定することはあまり行われていな
いのが現状である。Further, when attempting to analyze thousands of particle images with this microscope system, it is necessary to move the slide glass little by little to capture hundreds of images in order to change the field of view to be imaged. , Takes time and effort. For these reasons, it is the current situation that the size and shape of particles are not often measured from particle images for industrial powders.
【００１１】[0011]
【課題を解決するための手段】この発明は、粒子懸濁液
の流れをシース液で取り囲んだ流れに変換するシースフ
ローセルと、変換された懸濁液流に対して光を照射する
光照射手段と、照射された粒子を撮像する撮像手段と、
撮像された粒子像を解析する画像解析手段と、表示手段
とを備え、画像解析手段は、撮像された各粒子像の面積
および周囲長についての粒子データを測定し、その粒子
データから粒子の粒径と円形度を算出する算出手段と、
粒径による粒度頻度データに基づいてヒストグラムを作
成すると共に粒径と円形度とに対応する２つのパラメー
タによる２次元スキャッタグラムを作成して表示手段に
それぞれ表示する図表作成手段と、撮像された各粒子像
を格納する記憶手段と、記憶手段に格納された各粒子像
を表示手段に一括表示する粒子像呼出手段とからなるこ
とを特徴とする粒子画像分析装置を提供するものであ
る。The present invention is directed to a sheath flow cell for converting a flow of a particle suspension into a flow surrounded by a sheath liquid, and a light irradiation means for irradiating the converted suspension flow with light. And an imaging means for imaging the irradiated particles,
The image analysis means for analyzing the imaged particle image, and the display means, the image analysis means measures the particle data about the area and the perimeter of each imaged particle image, the particle data of the particle from the particle data. Calculation means for calculating the diameter and circularity,
A chart creating means for creating a histogram based on the particle size frequency data based on the particle size, creating a two-dimensional scattergram with two parameters corresponding to the particle size and the circularity, and displaying the two-dimensional scattergram on the display means, respectively. The present invention provides a particle image analysis device comprising a storage means for storing a particle image and a particle image calling means for collectively displaying each particle image stored in the storage means on a display means.
【００１２】この発明の装置の分析対象は、ファインセ
ラミックス、顔料、化粧品用パウダーのような無機物の
粉体および食品添加物のような有機物の粉体を含むもの
であり、予め染料や標識試薬によって染色処理された粒
子であってもよい。The object of analysis of the apparatus of the present invention includes fine ceramics, pigments, powders of inorganic substances such as powders for cosmetics and powders of organic substances such as food additives. The particles may be dyed.
【００１３】シースフローセルは、粒子を含む試料液、
すなわち、粒子懸濁液の流れをシース液で包んで流すこ
とにより流体力学的効果によって、細いあるいは偏平な
流れに変換することができるセルであり、これには、従
来公知のものを用いることができる。The sheath flow cell is a sample liquid containing particles,
That is, it is a cell that can be converted into a thin or flat flow by a hydrodynamic effect by wrapping the flow of a particle suspension in a sheath liquid and flowing it. For this, a conventionally known cell can be used. it can.
【００１４】なお、シースフローセルに供給されるシー
ス液については、粒子懸濁液の性質（粒子や溶媒の性
質）に対応してその種類を選択することが好ましい。光
照射手段には、パルス発光するストロボやレーザ光源を
用いることが好ましい。連続的に発光する光源を用いる
こともできるが、この場合には、撮像手段にシャッター
を設ける必要がある。撮像手段には、一般的な２次元画
像を撮像するビデオカメラを用いることができる。The type of the sheath liquid supplied to the sheath flow cell is preferably selected according to the properties of the particle suspension (the properties of particles and solvent). As the light irradiation means, it is preferable to use a strobe that emits pulsed light or a laser light source. A light source that emits light continuously can be used, but in this case, it is necessary to provide a shutter on the image pickup means. A video camera that captures a general two-dimensional image can be used as the imaging means.
【００１５】光照射手段と撮像手段とはシースフローセ
ルを挟んで配置され、シースフローセルにおいて粒子懸
濁液が偏平な流れに変換される場合、光照射手段は、粒
子懸濁流の偏平な一面に直交して光を照射し、撮像手段
はその光軸上に配置されることが好ましい。The light irradiation means and the imaging means are arranged with the sheath flow cell interposed therebetween, and when the particle suspension is converted into a flat flow in the sheath flow cell, the light irradiation means is orthogonal to one flat surface of the particle suspension flow. It is preferable that the image pickup means is arranged on the optical axis thereof.
【００１６】画像解析手段は、１／３０秒ごとの撮像画
面を実時間で処理できるパイプライン処理方式の画像処
理回路、ならびにＣＰＵ，ＲＯＭ，ＲＡＭおよびＩ／Ｏ
ポートからなるマイクロコンピュータを備えることが好
ましい。表示手段には、例えば、ＣＲＴや液晶ディスプ
レイを用いることができる。The image analysis means is an image processing circuit of a pipeline processing system capable of processing an image pickup screen every 1/30 seconds in real time, and CPU, ROM, RAM and I / O.
It is preferable to provide a microcomputer including a port. As the display means, for example, a CRT or a liquid crystal display can be used.
【００１７】[0017]
【作用】シースフローセルは、粒子懸濁液の流れをシー
ス液で取り囲み、細いあるいは偏平な流れに変換し、光
照射手段は、変換された懸濁液流に対して光を照射し、
撮像手段は、光照射された粒子を撮像する。画像解析手
段は、撮像された粒子像を解析して解析結果を粒子像と
共に表示手段に表示する。The sheath flow cell surrounds the particle suspension flow with the sheath liquid and converts it into a thin or flat flow, and the light irradiation means irradiates the converted suspension flow with light,
The imaging means images the particles irradiated with light. The image analysis means analyzes the captured particle image and displays the analysis result together with the particle image on the display means.
【００１８】つまり、画像解析手段においては、算出手
段が、撮像された各粒子像の面積および周囲長について
の粒子データを算出し、そのデータから粒径と円形度を
算出し、図表作成手段が、粒径による粒度頻度データに
基づいてヒストグラムを作成すると共に、粒径と円形度
とに対応する２つのパラメータによる２次元スキャッタ
グラムを作成してそれらを表示手段に表示する。That is, in the image analysis means, the calculation means calculates particle data on the area and perimeter of each imaged particle image, calculates the particle size and circularity from the data, and the chart creation means A histogram is created based on the particle size frequency data based on the particle size, and a two-dimensional scattergram based on two parameters corresponding to the particle size and circularity is created and displayed on the display means.
【００１９】一方、記憶手段は、撮像された各粒子像を
格納し、粒子像呼出手段は、記憶手段に格納された各粒
子像を表示手段に一括表示する。つまり、この粒子分析
装置では、撮像された各粒子像から粒子の大きさや周囲
長を求め、また、実際の粒子の形態や凝集状態を一括表
示される粒子像で確認することができる。On the other hand, the storage means stores the picked-up particle images, and the particle image recall means collectively displays the particle images stored in the storage means on the display means. That is, in this particle analyzer, the size and perimeter of the particles can be obtained from the imaged particle images, and the actual morphology and agglomeration state of the particles can be confirmed by the particle images displayed collectively.
【００２０】具体的には、粒子懸濁液を透明なフローセ
ルに導き、その懸濁液を細い又は偏平な流れにする。そ
の流れに対して光照射することによって、流れの中の粒
子をビデオカメラで撮像する。撮像された各粒子像の投
影面積と周囲長を算出し、次に粒径と円形度を算出す
る。さらに、粒径による粒度ヒストグラムおよび粒径と
円形度の２次元スキャッタグラムを作成する。この２次
元スキャッタグラムと実際の粒子像を評価、確認するこ
とによって、粒子の円形度や凝集度合いに関する情報を
得ることができる。Specifically, the particle suspension is introduced into a transparent flow cell, and the suspension is made into a thin or flat flow. The particles in the stream are imaged with a video camera by illuminating the stream. The projected area and perimeter of each imaged particle image are calculated, and then the particle size and circularity are calculated. Further, a grain size histogram based on grain size and a two-dimensional scattergram of grain size and circularity are created. By evaluating and confirming this two-dimensional scattergram and the actual particle image, it is possible to obtain information on the circularity and the degree of aggregation of particles.
【００２１】[0021]
【実施例】この発明のフロー方式粒子画像分析装置の例
を図１および図２に示す。これらの図において、まず粒
子懸濁液はダイヤフラムポンプ等の吸引手段（図示して
いない）によって吸引ピペット１から吸引され、サンプ
ルフィルター２を通りフローセル５の上部の試料チャー
ジングライン３へ引き込まれる。サンプルフィルター２
によって、懸濁液中の粗大な粒子やごみが取り除かれ、
流路の細い（狭い）フローセル５が詰まらないようにし
ている。またこのサンプルフィルタ２は、粗大な凝集塊
をほぐす効果も持っている。DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS An example of a flow type particle image analyzer of the present invention is shown in FIGS. In these figures, first, the particle suspension is sucked from a suction pipette 1 by a suction means (not shown) such as a diaphragm pump, drawn through the sample filter 2 and drawn into the sample charging line 3 above the flow cell 5. Sample filter 2
Removes coarse particles and debris from the suspension,
The flow cell 5 having a narrow (narrow) flow path is prevented from being clogged. The sample filter 2 also has the effect of loosening coarse aggregates.
【００２２】測定する粒子が半透明状の場合には、その
粒子に対して適当な染色を施すのが好ましい。図１には
図示していないが、装置内に染色液ボトルを設け、吸引
した試料をその染色液で染色するための反応チャンバー
を付加してもよい。When the particles to be measured are translucent, it is preferable to dye the particles appropriately. Although not shown in FIG. 1, a staining solution bottle may be provided in the apparatus and a reaction chamber for staining the aspirated sample with the staining solution may be added.
【００２３】チャージングライン３に引き込まれた粒子
懸濁液は、シースシリンジ４を動作させることによって
フローセル５に導かれ、サンプルノズル５ａの先端から
懸濁液が少しずつ押し出される。それと同時にシース液
もシース液ボトル６からシース液チャンバー７を介して
フローセル５に送り込まれ、粒子懸濁液はそのシース液
で取り囲まれ、図２に示すように、液体力学的に懸濁液
流は偏平に絞られてフローセル５の内を流れ、廃液チャ
ンバー１４を介して排出される。このように偏平に絞ら
れた懸濁液流に対して、ストロボ８からパルス光を１／
３０秒ごとに周期的に照射することによって、１／３０
秒ごとに粒子の静止画像が対物レンズ９を介してビデオ
カメラ１０で撮像される。The particle suspension drawn into the charging line 3 is guided to the flow cell 5 by operating the sheath syringe 4, and the suspension is gradually extruded from the tip of the sample nozzle 5a. At the same time, the sheath liquid is also sent from the sheath liquid bottle 6 into the flow cell 5 through the sheath liquid chamber 7, the particle suspension is surrounded by the sheath liquid, and as shown in FIG. Is squeezed into a flat state, flows through the flow cell 5, and is discharged through the waste liquid chamber 14. The pulse light from the strobe 8 is 1 /
By irradiating periodically every 30 seconds, 1/30
Every second, a still image of the particles is captured by the video camera 10 via the objective lens 9.
【００２４】粒子を懸濁する溶媒は粒子特性（粒径や比
重）に応じて最適なものを選べばよい。また、懸濁液の
流れを確実に偏平にあるいは細く絞り込むため懸濁液の
特性に応じて、例えば溶媒の粘度や比重に応じて、シー
ス液の粘度や比重を変更するのが好ましい。図１には図
示していないが、複数種類のシース液ボトルを設け、測
定する試料に応じて使用するシース液の種類を容易に切
り換えられるような機構を付加してもよい。The solvent for suspending the particles may be optimally selected according to the particle characteristics (particle size and specific gravity). Further, in order to reliably flatten or narrow the flow of the suspension, it is preferable to change the viscosity and the specific gravity of the sheath liquid according to the characteristics of the suspension, for example, the viscosity and the specific gravity of the solvent. Although not shown in FIG. 1, a plurality of types of sheath liquid bottles may be provided and a mechanism may be added so that the type of sheath liquid used can be easily switched according to the sample to be measured.
【００２５】懸濁液流の偏平な面をビデオカメラ１０で
撮像すれば、ビデオカメラ１０の撮像エリア全体に渡っ
て粒子像を捉えることができ、１回の撮像で多数の粒子
を撮像できる。また、撮像される粒子の重心とビデオカ
メラ１０の撮像面との距離をほぼ一定にすることができ
るので、粒子の大きさに関わらず常にピントの合った粒
子像が得られる。さらに、流体力学的な効果によって、
偏平な粒子や細長い粒子の向きが揃いやすく、粒子像を
解析して得られる特徴パラメータは、ばらつきが小さく
再現性が良い。If the flat surface of the suspension flow is imaged by the video camera 10, a particle image can be captured over the entire imaging area of the video camera 10, and a large number of particles can be imaged by one imaging. Further, since the distance between the center of gravity of the particles to be imaged and the image pickup surface of the video camera 10 can be made substantially constant, a focused particle image can always be obtained regardless of the size of the particles. Furthermore, due to the hydrodynamic effect,
The orientation of flat particles and elongated particles is easily aligned, and the characteristic parameters obtained by analyzing the particle image have small variations and good reproducibility.
【００２６】複数回のパルス光照射によって撮像される
粒子像の数は、懸濁液流を偏平にした場合には、ビデオ
カメラ１０の撮像エリアの面積、試料流の厚み、粒子懸
濁液の単位体積当たりの粒子数、および撮像回数（フレ
ーム数）によって決まる。例えば、撮像エリアを２００
×２００μｍ、試料流の厚みを５μｍ、粒子濃度を１０
０００個／μｌ、撮像フレーム数を１８００（撮像時間
を６０秒）とした時に撮像される粒子数は３６００個と
なる。When the suspension flow is flattened, the number of particle images imaged by plural times of pulsed light irradiation is the area of the imaging area of the video camera 10, the thickness of the sample flow, and the particle suspension. It depends on the number of particles per unit volume and the number of times of imaging (the number of frames). For example, if the imaging area is 200
× 200 μm, sample flow thickness 5 μm, particle concentration 10
When the number of particles is 000 / μl and the number of imaging frames is 1800 (imaging time is 60 seconds), the number of particles imaged is 3600.
【００２７】撮像エリアの面積は、ビデオカメラ１０の
受光面に対する結像倍率とそのサイズによって決まる。
対物レンズ９の倍率を大きくすれば撮像エリアが小さく
なるが、小さな粒子まで大きく撮像できる。対物レンズ
９の倍率を小さくすれば撮像エリアが大きくなり、大き
な粒子を撮像するのに適している。この装置では、対物
レンズ９の倍率を選択あるいは測定途中に切り換えでき
るようにしており（図示していない）、粒径の測定レン
ジを広くしている。The area of the image pickup area is determined by the image forming magnification and the size of the light receiving surface of the video camera 10.
If the magnification of the objective lens 9 is increased, the imaging area is reduced, but even small particles can be imaged large. If the magnification of the objective lens 9 is reduced, the imaging area becomes large, which is suitable for capturing large particles. In this device, the magnification of the objective lens 9 can be selected or switched during measurement (not shown), and the measurement range of particle size is widened.
【００２８】ビデオカメラ１０からの画像信号は、画像
処理装置１１で処理され、モニターテレビ１２に表示さ
れる。１３は各種の操作等を行うためのキーボード（又
はマウス）である。The image signal from the video camera 10 is processed by the image processing device 11 and displayed on the monitor television 12. A keyboard (or mouse) 13 is used to perform various operations.
【００２９】１／３０秒ごとの粒子撮像画面に対する画
像処理の手順を図５に示す。画像信号は、画像処理装置
１１に取り込まれてＡ／Ｄ変換され、画像データとして
取り込まれる（ステップＳ１）。まず懸濁液流に対する
照射光の強度むら（シェーディング）を補正するための
バックグランド補正が行われる（ステップＳ２）。FIG. 5 shows a procedure of image processing for the particle image pickup screen every 1/30 seconds. The image signal is captured by the image processing device 11, A / D converted, and captured as image data (step S1). First, background correction is performed to correct the intensity unevenness (shading) of the irradiation light with respect to the suspension flow (step S2).
【００３０】具体的には、粒子がフローセル５を通過し
ていない時に光照射して得られる画像データを、測定前
にあらかじめ取り込んでおき、その画像データと実際の
粒子撮像画面の画像データとを比較演算することであ
り、画像処理として一般的によく知られた処理である。
次に、粒子像の輪郭を的確に抽出するための前処理とし
て輪郭強調処理を行う（ステップＳ３）。具体的には、
一般的によく知られたラプラシアン強調処理を行う。Specifically, the image data obtained by irradiating light when the particles do not pass through the flow cell 5 are captured in advance before measurement, and the image data and the image data of the actual particle imaging screen are collected. It is a comparison calculation, which is generally well known as image processing.
Next, a contour enhancement process is performed as a pre-process for accurately extracting the contour of the particle image (step S3). In particular,
Generally well-known Laplacian emphasis processing is performed.
【００３１】次に、画像データをある適当なスレシホー
ルドレベルで２値化する（ステップＳ４）。次に、２値
化された粒子像に対してエッジ点かどうかを判定すると
ともに、着目しているエッジ点に対して隣合うエッジ点
がどの方向にあるかの情報、すなわちチェインコードを
生成する（ステップＳ５）。次に、このチェインコード
を参照しながら粒子像のエッジトレースを行い、各粒子
像の総画素数、総エッジ数、斜めエッジ数を求める（ス
テップＳ６）。Next, the image data is binarized at a certain appropriate threshold level (step S4). Next, it is determined whether or not the binarized particle image is an edge point, and information indicating in which direction the adjacent edge point is with respect to the focused edge point, that is, a chain code is generated. (Step S5). Next, edge tracing of the particle image is performed with reference to this chain code, and the total number of pixels, the total number of edges, and the number of diagonal edges of each particle image are obtained (step S6).
【００３２】高性能のパイプライン処理可能な画像処理
装置を使用すれば、以上の画像処理を、１／３０秒ごと
に撮像される画面に対してリアルタイムに処理すること
ができる。この装置では、ある倍率で撮像される複数の
画面に対して上記画像処理を繰り返し行い、次に異なる
撮像倍率に切り換えて撮像し、同様の画像処理を行う。
また、撮像されたフレームから粒子像の切り出しを行
い、切り出した粒子像を画像処理装置１１の画像メモリ
に格納する（ステップＳ７）。If an image processing apparatus capable of high-performance pipeline processing is used, the above image processing can be performed in real time on a screen imaged every 1/30 seconds. In this apparatus, the above-mentioned image processing is repeatedly performed on a plurality of screens imaged at a certain magnification, and then the imaging is performed by switching to a different imaging magnification, and the same image processing is performed.
Further, the particle image is cut out from the captured frame, and the cut out particle image is stored in the image memory of the image processing apparatus 11 (step S7).
【００３３】撮像が終了すると（ステップＳ８）、各粒
子像に対して求められた総画素数、総エッジ数、斜めエ
ッジ数から、まず下記の式によって各粒子像の投影面積
Ｓと周囲長Ｌを求める。When the image pickup is completed (step S8), the projected area S and the perimeter L of each particle image are first calculated by the following formulas from the total number of pixels, the total number of edges, and the number of oblique edges obtained for each particle image. Ask for.
【００３４】図１０に示すように、２値画像の周囲のエ
ッジの中心を結んでできる枠内の面積Ｓおよび枠の長さ
（周期長Ｌ）は、１画素当たりの面積を１とした場合、
面積Ｓ＝総画素数−（総エッジ数×０．５）−１……(1)
周囲長Ｌ＝（総エッジ数−斜めエッジ数）＋（斜めエッジ数×√２）……(2）As shown in FIG. 10, the area S within the frame formed by connecting the centers of the edges around the binary image and the length of the frame (cycle length L) are when the area per pixel is 1. , Area S = total number of pixels− (total number of edges × 0.5) −1 (1) Perimeter L = (total number of edges−number of diagonal edges) + (number of diagonal edges × √2) …… (2 )
【００３５】次に、上記面積Ｓと周囲長Ｌを用いて円相
当径と円形度を求める（ステップＳ９）。厳密に言う
と、縦横線と斜め４５°の線で粒子像の輪郭を表わす
と、上式で求められる周囲長Ｌは丸い粒子像の場合、
１．０５倍程度長くなり、円形度を求める際に少し補正
が必要となる。円形当径とは、粒子像の投影面積と同じ
面積を持つ円を想定し、その円の直径のことであり、式
（３）で表される。円形度とは、例えば式（４）で定義
される値であり、粒子像が円形の時に円形度は１にな
り、粒子像が細長くなればなるほど円形度は小さい値に
なる。Next, using the area S and the perimeter L, the equivalent circle diameter and circularity are obtained (step S9). Strictly speaking, if the contour of the particle image is represented by vertical and horizontal lines and a line of 45 ° diagonally, the perimeter L obtained by the above equation is
It is about 1.05 times longer, and some correction is required when obtaining the circularity. The circular equivalent diameter is the diameter of the circle assuming a circle having the same area as the projected area of the particle image, and is represented by equation (3). The circularity is a value defined by, for example, Expression (4), and the circularity becomes 1 when the particle image is circular, and the circularity becomes smaller as the particle image becomes elongated.
【００３６】 円相当径＝（粒子投影像面積値／π）1/2×２……(3) 円形度＝（粒子像と同じ投影面積値を持つ円の周囲長）／粒子投影像の周囲長 ）……(4)Equivalent diameter of circle = (particle projection image area value / π) 1/2 × 2 (3) Circularity = (perimeter of circle having projection area value same as particle image) / perimeter of particle projection image Long) …… (4)
【００３７】各粒子像の円相当径が求められれば、次に
その値をもとにして粒度頻度データを作成する（ステッ
プＳ１０）。工業用の粉体は多種多様で粒径も非常に広
い範囲に渡っている。従って、一般的に粒径はＬＯＧ
（対数）変換し、ＬＯＧ変換した値を等分割した上で粒
度頻度データを求める。When the equivalent circle diameter of each particle image is obtained, the particle size frequency data is created based on the value (step S10). There are a wide variety of industrial powders and their particle sizes are very wide. Therefore, the particle size is generally LOG
The (logarithmic) conversion is performed, and the LOG-converted value is equally divided, and then the granularity frequency data is obtained.
【００３８】ところで、粒子撮像画面（フレーム）にお
いて、画面の端にかかる粒子像からは正しくその粒子の
円相当径や円形度を求めることはできない。従って、画
面の端にかかって写っている粒子像は無視する必要があ
る。図３に示すように、大きな粒子像ほど画面の端にか
かる確率が高いことは明らかであり、画像処理法で粒度
分布を正しく求めるにはこのことを考慮しなければなら
ない。そして、粒子像の大きさに応じてその頻度値を補
正する。By the way, on the particle image pickup screen (frame), the equivalent circle diameter and the circularity of the particle cannot be correctly obtained from the particle image on the edge of the screen. Therefore, it is necessary to ignore the particle image that appears on the edge of the screen. As shown in FIG. 3, it is clear that the larger the particle image, the higher the probability of hitting the edge of the screen, and this must be taken into consideration in order to correctly obtain the particle size distribution by the image processing method. Then, the frequency value is corrected according to the size of the particle image.
【００３９】ビデオカメラ１０の撮像エリアに対して粒
子像が十分小さい場合には、画面の端にかからない粒子
像の重心の存在エリアは、ほぼ撮像エリアと同じであ
る。粒子像が大きい場合ほど、画面の端にかからない粒
子像の重心の存在エリアは、撮像エリアに対して大きく
狭まる。When the particle image is sufficiently smaller than the image pickup area of the video camera 10, the area where the center of gravity of the particle image does not reach the edge of the screen is almost the same as the image pickup area. As the particle image is larger, the area where the center of gravity of the particle image that does not reach the edge of the screen is narrower than the imaging area.
【００４０】すなわち、大きな粒子ほど実質の試料分析
量が減ることになり、大きな粒子ほど相対的に頻度が小
さくなる。試料分析量は、画面の端にかからない粒子像
の重心の存在エリアの面積に比例する。従って、円相当
径がｄ〜（ｄ＋Δｄ）の粒子頻度データは、式（５）で
補正すればよい。つまり、頻度補正係数は、
（ビデオカメラの撮像エリアの面積）／｛（撮像エリアＸ方向サイズ-d）×（
撮像エリアＹ方向サイズ-d）｝……(5)
となる。ビデオカメラの撮像エリアと画像処理対象エリ
アが異なる場合には、上記撮像エリアを画像処理対象エ
リアに置き換えて算出する。That is, the larger the particles, the smaller the actual sample analysis amount, and the larger the particles, the smaller the frequency. The sample analysis amount is proportional to the area of the area where the center of gravity of the particle image does not lie on the edge of the screen. Therefore, the particle frequency data for which the equivalent circle diameter is d to (d + Δd) may be corrected by the equation (5). That is, the frequency correction coefficient is (area of image pickup area of video camera) / {(image pickup area X direction size-d) × (image pickup area Y direction size-d)} (5). When the image pickup area of the video camera is different from the image processing target area, the image pickup area is replaced with the image processing target area for calculation.
【００４１】粒度頻度データは、まずそれぞれの撮像倍
率で撮像された粒子像に対して独立に求める。それぞれ
の撮像倍率での粒径測定範囲は異なり、例えば図６に示
すように、高倍率撮像での粒径測定範囲を１〜３０μ
ｍ、低倍率撮像での粒径測定範囲を１５〜３００μｍと
している。この例では、１５〜３０μｍの範囲をオーバ
ーラップさせている。The particle size frequency data is first obtained independently for the particle images taken at the respective imaging magnifications. The particle size measurement range at each imaging magnification is different. For example, as shown in FIG. 6, the particle size measurement range at high magnification imaging is 1 to 30 μm.
m, and the particle size measurement range in low-magnification imaging is 15 to 300 μm. In this example, the range of 15 to 30 μm is overlapped.
【００４２】図６の例は、粒径が１５〜３０μｍの範囲
を越えて大きくばらついている粒子の例であり、高倍率
撮像と低倍率撮像でのそれぞれの粒度頻度データをつな
ぎ合わせる必要がある。そのためには、まずそれぞれの
撮像倍率での試料分析量の比に応じて、次式のような頻
度補正を行う必要がある。低撮像倍率では撮像エリアが
広いので、一般的に試料分析量を多くすることができ
る。
（高倍率撮像での頻度値）×（低倍率撮像での試料分析
量）／（高倍率撮像での試料分析量）
なお、試料分析量は、
（撮像面積）×（粒子懸濁液流の厚み）×（撮像フレー
ム数）
で求めることができる。The example of FIG. 6 is an example of particles having a large variation in particle size exceeding the range of 15 to 30 μm, and it is necessary to connect the particle size frequency data for high-magnification imaging and that for low-magnification imaging. . For that purpose, first, it is necessary to perform frequency correction as shown in the following formula according to the ratio of the sample analysis amount at each imaging magnification. Since the imaging area is wide at a low imaging magnification, the sample analysis amount can generally be increased. (Frequency value in high-magnification imaging) x (Sample analysis amount in low-magnification imaging) / (Sample analysis amount in high-magnification imaging) The sample analysis amount is (imaging area) x (particle suspension flow It can be calculated by (thickness) × (number of imaging frames).
【００４３】上記のような試料分析量の違いによる頻度
補正を行っても、必ずしもそれぞれの頻度データによる
頻度分布曲線が滑らかにつながらず、つなぎめで段差が
生じることがある。その最も大きな原因は、粒子懸濁液
の粒子濃度が薄い場合に、撮像された粒子数が少なく
て、図６の破線で示すように頻度分布曲線が大きくがた
つく場合である。Even if the frequency correction is performed according to the difference in the sample analysis amount as described above, the frequency distribution curves based on the respective frequency data are not always connected smoothly, and a step may occur at the connection. The largest cause thereof is that when the particle concentration of the particle suspension is low, the number of imaged particles is small and the frequency distribution curve is largely distorted as shown by the broken line in FIG.
【００４４】他の原因として、対物レンズあるいは投影
レンズの倍率が仕様通りの値になっていないために、高
倍率撮像と低倍率撮像での真の試料分析量が予測とは異
なり、上記試料分析量の違いによる頻度補正が正確でな
くなる場合である。ただし、この撮像倍率が不正確であ
ることによる段差の原因は、あらかじめ測定装置１台ご
とに撮像倍率を校正することによって解決することがで
きる。 この装置では、異なる撮像倍率での粒径測定範
囲を一部オーバーラップさせ、そのオーバーラップ測定
範囲において、それぞれの撮像倍率での頻度値を加重平
均するようにしている。As another cause, since the magnification of the objective lens or the projection lens is not the value as specified, the true sample analysis amount in high-magnification imaging and low-magnification imaging is different from the prediction, and the above sample analysis is different. This is the case where the frequency correction due to the difference in quantity becomes inaccurate. However, the cause of the step due to the inaccurate imaging magnification can be solved by calibrating the imaging magnification for each measuring device in advance. In this apparatus, the particle size measurement ranges at different imaging magnifications are partially overlapped, and the frequency values at the respective imaging magnifications are weighted averaged in the overlapping measurement range.
【００４５】オーバーラップ測定範囲の上限に近いほど
低倍率撮像での頻度値に大きな重みを付け、下限に近い
ほど高倍率撮像での頻度値に大きな重みを付けて加重平
均する。このような加重平均法による頻度補正をするこ
とによって、撮像粒子数が少ない場合でも、異なる撮像
倍率での頻度分布データを滑らかにつなぎ合わせること
ができる。例えば、高倍率撮像と低倍率撮像でのオーバ
ーラップ範囲を１５〜３０μｍとした場合、そのオーバ
ーラップ範囲内の粒径ｄ〜ｄ＋Δｄ（μｍ）の粒子頻度
値ｆ（ｄ）は、次式で算出する。The closer the value to the upper limit of the overlap measurement range, the greater the weight to the frequency value in low-magnification imaging, and the closer to the lower limit to the frequency value in high-magnification imaging, the greater the weight, and the weighted average. By performing frequency correction by such a weighted average method, frequency distribution data at different imaging magnifications can be smoothly joined together even when the number of imaged particles is small. For example, when the overlap range in high-magnification imaging and low-magnification imaging is 15 to 30 μm, the particle frequency value f (d) of the particle diameters d to d + Δd (μm) within the overlap range is calculated by the following formula. To do.
【００４６】ｆ（ｄ）＝高倍率撮像頻度値（ｄ）×（１
−（ｄ−15）／（30−15））＋低倍率撮像頻度値（ｄ）
×（１−（30−ｄ）／（30−15））
以上のようにして求められた粒度頻度データを用いて、
さらに累積粒度データを求める。例えば、個数基準の累
積粒度データ（％）は、次式で算出する。
粒度ｄにおける累積粒度（ｄ）＝（粒径ｄ以下の粒子
数）×１００／（全粒子数）F (d) = high-magnification imaging frequency value (d) × (1
-(D-15) / (30-15)) + low magnification imaging frequency value (d)
× (1- (30-d) / (30-15)) Using the particle size frequency data obtained as described above,
Further, the cumulative grain size data is obtained. For example, the number-based cumulative particle size data (%) is calculated by the following formula. Cumulative particle size (d) at particle size d = (number of particles of particle size d or less) x 100 / (total number of particles)
【００４７】次に、円相当径と円形度の２つのパラメー
タによる２次元スキャッタ頻度データを求める（ステッ
プＳ１１）。この場合にも、まず高倍率撮像と低倍率撮
像のそれぞれに対して２次元頻度データを求める。次
に、粒度頻度データの補正処理と同様に、粒子像の大き
さの違いによる頻度補正、異なる撮像倍率での試料分析
量の違いによる頻度補正を行う。さらに、異なる撮像倍
率でのオーバーラップ測定範囲での２次元頻度補正を、
前記粒度頻度データのつなぎ合わせの時と同様に行う。Next, the two-dimensional scatter frequency data based on the two parameters of the circle equivalent diameter and the circularity are obtained (step S11). Also in this case, first, two-dimensional frequency data is obtained for each of high-magnification imaging and low-magnification imaging. Next, similar to the correction processing of the particle size frequency data, the frequency correction is performed due to the difference in the size of the particle image, and the frequency correction is performed due to the difference in the sample analysis amount at different imaging magnifications. In addition, two-dimensional frequency correction in the overlap measurement range at different imaging magnification,
The same operation is performed as when the grain size frequency data is connected.
【００４８】上記のようにして求められた粒度頻度デー
タ、累積粒度データ、および円相当径と円形度の２次元
頻度データを用いて、さらに平均粒径、粒径の標準偏
差、モード径、１０％径、５０％径、９０％径、平均円
形度、円形度標準偏差等を算出する（ステップＳ１
２）。Using the particle size frequency data, the cumulative particle size data, and the two-dimensional frequency data of the equivalent circle diameter and the circularity obtained as described above, the average particle diameter, the standard deviation of the particle diameter, the mode diameter, 10 % Diameter, 50% diameter, 90% diameter, average circularity, circularity standard deviation, etc. are calculated (step S1).
2).
【００４９】モード径とは、粒度頻度値が最大であると
ころの粒径のことを指す。１０％径、５０％径、９０％
径は、累積粒度データの値がそれぞれ１０、５０、９０
％の値になるところの粒径のことを指す。すなわち、５
０％径とは、粒径の中心値のことであり、メジアン径と
も言う。The mode diameter refers to the particle size at which the particle size frequency value is maximum. 10% diameter, 50% diameter, 90%
As for the diameter, the values of the cumulative particle size data are 10, 50 and 90, respectively.
It means the particle size at which the value of% is reached. That is, 5
The 0% diameter is the center value of the particle diameter, and is also called the median diameter.
【００５０】以上のようにして求められた頻度データお
よび解析結果から、図７、図８に示すような粒度ヒスト
グラム、円相当径と円形度の２次元スキャッタグラム、
および平均粒径や５０％径等の解析結果を表示する（ス
テップＳ１３）。図７では、横軸をＬＯＧ変換した円相
当径、縦軸を頻度％と累積％の２つの意味に割当て、累
積粒度分布曲線の表示も同時に表示している。図８に示
すスキャッタグラム表示では、横軸をＬＯＧ変換した円
相当径、縦軸を円形度としており、各分割点（ドット）
の色を２次元頻度値に応じて変えるようにしている。From the frequency data obtained as described above and the analysis results, a grain size histogram as shown in FIGS. 7 and 8, a two-dimensional scattergram of circle equivalent diameter and circularity,
And the analysis results such as the average particle diameter and the 50% diameter are displayed (step S13). In FIG. 7, the horizontal axis is LOG-converted equivalent circle diameter, and the vertical axis is assigned to two meanings of frequency% and cumulative%, and the cumulative particle size distribution curve is also displayed at the same time. In the scattergram display shown in FIG. 8, the horizontal axis represents the LOG-converted circle equivalent diameter and the vertical axis represents the circularity, and each division point (dot)
The color of is changed according to the two-dimensional frequency value.
【００５１】この装置では、上記のように撮像した粒子
像から円相当径や円形度を求めるだけでなく、撮像した
粒子像を記憶しておき、測定後に大きさ別にクラス分け
して図４に示すように、一括表示する機能も有してい
る。もっとも、画像を記憶する画像メモリの容量に制限
があるので、撮像された全ての粒子像を記憶、表示する
わけではない。撮像された粒子像を一括表示できる機能
を有しているので、粒子の形態や凝集状態を直接使用者
が確認することができる。In this apparatus, not only the equivalent circle diameter and the circularity are obtained from the imaged particle image as described above, but also the imaged particle image is stored, and after the measurement, the particles are classified according to size and shown in FIG. As shown, it also has the function of displaying all at once. However, since the capacity of the image memory that stores images is limited, not all the captured particle images are stored and displayed. Since it has a function of displaying the imaged particle images all at once, the user can directly check the morphology and agglomeration state of the particles.
【００５２】粒子どうし凝集することが重要な意味を持
つような場合には、図４に示す各枠内の粒子像につい
て、一次（単独）粒子像か、２個凝集粒子像か、３個凝
集粒子像か、高次凝集塊か、あるいは対象外の粒子か
を、使用者が指定しキーボード１３を用いて入力する。
その指定結果をもとにして、凝集している粒子の数の比
率を自動的に計算することができる。もし、一括表示さ
れた粒子像の中に凝集粒子像が全く無い場合には、上記
２次元スキャッタグラムでの円形度算出値は、真に粒子
の円形度を表していると考えてよい。When it is important to agglomerate particles, the particle images in each frame shown in FIG. 4 are primary (single) particle images, two agglomerated particle images, or three agglomerated particles. The user designates a particle image, a higher-order aggregate, or a non-target particle and inputs it using the keyboard 13.
Based on the designated result, the ratio of the number of agglomerated particles can be automatically calculated. If there is no aggregated particle image in the particle images displayed collectively, it can be considered that the calculated circularity value in the two-dimensional scattergram truly represents the circularity of particles.
【００５３】また、上記指定結果をもとにして、一次粒
子像だけを対象にして画像解析し直すこともできる。記
憶できる粒子像の数に限りがあるので、再現性の良い解
析結果が得られない場合もあるが、対象外の粒子（ごみ
等）や凝集粒子を除いて解析するので、より正確な粒度
分布、円形度が求められる。It is also possible to re-analyze only the primary particle image based on the above designated result. Since the number of particle images that can be stored is limited, it may not be possible to obtain reproducible analysis results. However, since particles other than the target particles (such as dust) and agglomerated particles are analyzed, a more accurate particle size distribution can be obtained. , Circularity is required.
【００５４】また、一括表示された粒子像により、使用
者が、測定した粒子が球形であることを確認した場合に
は、凝集しやすい粒子でも円形度が１に近い粒子だけに
限定して粒度解析しなおせば、より正確な粒度分布が求
められる。また、粒子が球形である場合には、図９に示
すように、円相当径と円形度による２次元スキャッタグ
ラムにおいて、凝集粒子が分布していると考えられる領
域を推定することもできる。In addition, when the user confirms that the measured particles are spherical by the particle images displayed collectively, the particle size is limited to particles having a circularity close to 1 even if the particles easily aggregate. A more accurate particle size distribution can be obtained by re-analyzing. Further, when the particles are spherical, as shown in FIG. 9, it is possible to estimate a region in which the agglomerated particles are considered to be distributed in the two-dimensional scattergram based on the circle equivalent diameter and the circularity.
【００５５】図９の例では、点線で囲んだ枠内に分布す
る粒子は、凝集粒子と推定している。粒径の揃っている
球形の粒子が２個凝集あるいは３個凝集した場合の粒子
像では、その投影面積は大きく、円相当径は約√２倍あ
るいは√３倍になり、円形度は０．９以下と小さくな
る。点線で囲った枠内の粒子数を計算すれば、粒子凝集
度合いに関する指標を求めることができる。In the example of FIG. 9, the particles distributed in the frame surrounded by the dotted line are presumed to be agglomerated particles. In a particle image in which two or three spherical particles having a uniform particle size are aggregated, the projected area is large, the equivalent circle diameter is about √2 times or √3 times, and the circularity is 0. It becomes as small as 9 or less. By calculating the number of particles in the frame surrounded by the dotted line, an index relating to the degree of particle aggregation can be obtained.
【００５６】この装置では、図９の例のように円相当径
と円形度による２次元スキャッタグラムにおいてある２
次元領域を設定し、その領域内あるいは領域外の粒子の
データだけに限定して粒度解析、円形度解析させること
もできる。このような機能を利用することによって、ご
みや凝集粒子を除いての粒度分布や平均円形度を求め
る、あるいは凝集している粒子数の比率を推定するとい
ったことが可能になる。このような２次元領域は、測定
する試料の種類ごとに、使用者がキーボード１３やマウ
スを使って任意に設定、変更できる。In this apparatus, as in the example of FIG. 9, the two-dimensional scattergram based on the equivalent circle diameter and the circularity is 2
It is also possible to set a dimensional region and limit particle size analysis and circularity analysis to data of particles within or outside the region. By using such a function, it becomes possible to obtain the particle size distribution and average circularity excluding dust and agglomerated particles, or to estimate the ratio of the number of agglomerated particles. Such a two-dimensional area can be arbitrarily set and changed by the user using the keyboard 13 or the mouse for each type of sample to be measured.
【００５７】以上のように、このフロー方式粒子画像分
析装置では、粒子像を使用者が直接目で確認できること
はもちろんのこと、従来の電気的検知帯法やレーザ解析
散乱法の測定装置では得られなかった定量的な情報、す
なわち円形度や凝集度合い等の新規の情報が得られる。
また、円相当径と円形度の２次元スキャッタグラムによ
って、試料中のごみや凝集粒子の分布領域を推定するこ
とができ、その領域内のデータを除外して粒度解析すれ
ば、より正確な粒度分布が求められる。As described above, in this flow type particle image analyzer, not only the user can directly check the particle image with the eyes, but also the conventional measuring device using the electric detection zone method or the laser analysis scattering method can obtain the particle image. Quantitative information that could not be obtained, that is, new information such as circularity and degree of aggregation is obtained.
In addition, the distribution area of dust and agglomerated particles in the sample can be estimated by the two-dimensional scattergram of the equivalent circle diameter and the circularity. If the data in that area is excluded and particle size analysis is performed, a more accurate particle size can be obtained. Distribution is required.
【００５８】[0058]
【発明の効果】この発明は、次のような効果を奏する。
１．粒子像を画像解析することによって、粒子の大きさ
（円相当径）だけでなく、粒子像の周囲長や円形度の情
報も求められる。
２．粒子懸濁液の流れを、シース液によって流体力学的
に細い又は偏平な流れにするので、粒子の大きさに関わ
らず、粒子の重心の通過する位置は、撮像方向に対して
ほとんど変動しないため、常にピントの合った粒子像が
撮像され、従来の顕微鏡画像処理法より信頼性の高い測
定結果が得られる。The present invention has the following effects. 1. By performing image analysis on the particle image, not only the size of the particle (equivalent circle diameter), but also information about the perimeter and circularity of the particle image can be obtained. 2. Since the flow of the particle suspension is made hydrodynamically thin or flat by the sheath liquid, the position where the center of gravity of the particle passes does not change much with respect to the imaging direction regardless of the size of the particle. , Particle images that are in focus are always captured, and more reliable measurement results can be obtained than with conventional microscope image processing methods.
【００５９】３．粒子懸濁液流を流体力学的に細い又は
偏平な試料流にするので、偏平な粒子や細長い粒子の向
きが揃いやすく、粒子像から求められる円相当径や円形
度のばらつきは小さく再現性が良い。
４．撮像、記憶した粒子像を、測定後に表示手段に一括
表示できるので、粒子の形態や凝集状態を容易に確認す
ることができる。3. Since the particle suspension flow is hydrodynamically made into a thin or flat sample flow, the orientation of flat particles and elongated particles can be easily aligned, and the variations in equivalent circle diameter and circularity required from particle images are small and reproducibility is small. good. 4. Since the imaged and stored particle images can be collectively displayed on the display means after the measurement, it is possible to easily confirm the morphology and agglomeration state of the particles.
【００６０】５．撮像、記憶した粒子像により、粒子が
凝集しているかどうかを使用者が目視で識別分類でき、
凝集している粒子の比率を求めることができる。
６．粒子像一括表示により粒子が球形であることが確認
された場合には、凝集しやすい粒子でも、円形度が１に
近い粒子のデータだけに限定して粒度解析すれば、より
正確な粒度分布が求められる。
７．粒子像一括表示により粒子が球形であることが確認
された場合には、円相当径と円形度の２次元スキャッタ
グラムから粒子凝集度合いに関する指標を得ることがで
きる。5. The captured and stored particle image allows the user to visually identify and classify whether the particles are agglomerated,
The ratio of particles that are agglomerated can be determined. 6. If it is confirmed by the particle image batch display that the particles are spherical, a more accurate particle size distribution can be obtained if particle size analysis is performed by limiting the data to those particles that tend to agglomerate and whose circularity is close to 1. Desired. 7. When it is confirmed by the particle image collective display that the particles are spherical, an index relating to the degree of particle aggregation can be obtained from the two-dimensional scattergram of the equivalent circle diameter and the circularity.
【図面の簡単な説明】[Brief description of drawings]
【図１】実施例の構成説明図である。FIG. 1 is a diagram illustrating a configuration of an embodiment.
【図２】図１の要部拡大断面図である。FIG. 2 is an enlarged cross-sectional view of a main part of FIG.
【図３】粒子撮像画面の例を示す説明図である。FIG. 3 is an explanatory diagram showing an example of a particle imaging screen.
【図４】モニターテレビの表示画像の例を示す説明図で
ある。FIG. 4 is an explanatory diagram showing an example of a display image on a monitor television.
【図５】実施例の処理手順を示すフローチャートであ
る。FIG. 5 is a flowchart showing a processing procedure of the embodiment.
【図６】撮像倍率の異なるデータから合成したヒストグ
ラムである。FIG. 6 is a histogram synthesized from data having different imaging magnifications.
【図７】粒度分布および累積粒度分布の表示例を示す説
明図である。FIG. 7 is an explanatory diagram showing a display example of a particle size distribution and a cumulative particle size distribution.
【図８】スキャッタグラムの表示例を示す説明図であ
る。FIG. 8 is an explanatory diagram showing a display example of a scattergram.
【図９】スキャッタグラムにおける領域設定例を示す説
明図である。FIG. 9 is an explanatory diagram showing an example of area settings in a scattergram.
【図１０】粒子の撮影面積と周囲長の算出を示す説明図
である。FIG. 10 is an explanatory diagram showing calculation of an imaging area of particles and a perimeter.
１ 吸引ピペット ２ サンプルフィルター ３ 試料チャージングライン ４ シースシリンジ ５ フローセル ６ シース液ボトル ７ シース液チャンバー ８ ストロボ ９ 対物レンズ １０ ビデオカメラ １１ 画像処理装置 １２ モニターテレビ １３ キーボード 1 suction pipette 2 sample filter 3 Sample charging line 4 sheath syringe 5 flow cells 6 sheath liquid bottle 7 Sheath liquid chamber 8 strobes 9 Objective lens 10 video cameras 11 Image processing device 12 monitor TV 13 keyboard
───────────────────────────────────────────────────── フロントページの続き (58)調査した分野(Int.Cl.7，ＤＢ名) G01N 15/14 G01N 15/02 ─────────────────────────────────────────────────── ─── Continuation of the front page (58) Fields surveyed (Int.Cl. 7 , DB name) G01N 15/14 G01N 15/02
Claims (9)
だ流れに変換するシースフローセルと、変換された懸濁
液流に対して光を照射する光照射手段と、照射された粒
子を撮像する撮像手段と、撮像された粒子像を解析する
画像解析手段と、表示手段とを備え、画像解析手段は、
撮像された各粒子像の面積および周囲長についての粒子
データを測定し、その粒子データから粒子の粒径と円形
度を算出する算出手段と、粒径による粒度頻度データに
基づいてヒストグラムを作成すると共に粒径と円形度と
に対応する２つのパラメータによる２次元スキャッタグ
ラムを作成して表示手段にそれぞれ表示する図表作成手
段と、撮像された各粒子像を格納する記憶手段と、記憶
手段に格納された各粒子像を表示手段に一括表示する粒
子像呼出手段とからなることを特徴とする粒子画像分析
装置。1. A sheath flow cell for converting a flow of a particle suspension into a flow surrounded by a sheath liquid, a light irradiation means for irradiating the converted suspension flow with light, and an image of the irradiated particle. Image capturing means, image analyzing means for analyzing the captured particle image, and display means, and the image analyzing means comprises:
Particle data for the area and perimeter of each imaged particle is measured, and a histogram is created based on the particle size data and the calculation means for calculating the particle size and circularity of the particle from the particle data. In addition, a chart creating means for creating a two-dimensional scattergram by two parameters corresponding to the particle size and the circularity and displaying it on the display means, a storage means for storing each imaged particle image, and a storage means A particle image analyzing device comprising: a particle image recalling unit that collectively displays the generated particle images on a display unit.
ら円形度頻度データおよび／又は円形度の平均値と標準
偏差を算出して表示手段に表示する演算手段をさらに備
えてなる請求項１記載の粒子画像分析装置。2. The image analysis means further comprises calculation means for calculating circularity frequency data and / or average value and standard deviation of circularity from the circularity of individual particles and displaying the average and standard deviation on the display means. 1. The particle image analyzer according to 1.
種類を選択し、シースフローセルに供給する供給手段お
よび／又は粒子を予め染色するための染色手段をさらに
備えてなる請求項１記載の粒子画像分析装置。3. The method according to claim 1, further comprising a supply means for selecting the type of the sheath liquid according to the characteristics of the particle suspension and supplying it to the sheath flow cell and / or a dyeing means for dyeing the particles in advance. The described particle image analyzer.
な流れに変換すると共に、撮像手段が粒子懸濁液流の偏
平な面を撮像することを特徴とする請求項１記載の粒子
画像分析装置。4. The particle image analysis according to claim 1, wherein the sheath flow cell converts the particle suspension into a flat flow, and the imaging means images a flat surface of the particle suspension flow. apparatus.
段から得られる撮像画面について、撮像画面の端にかか
っている粒子像を無視し、粒度頻度データを円相当径の
大きさに応じて補正することを特徴とする請求項１記載
の粒子画像分析装置。5. The calculation means and the chart creation means ignore the particle image on the edge of the image pickup screen obtained from the image pickup means, and correct the particle size frequency data according to the size of the circle equivalent diameter. The particle image analysis device according to claim 1, wherein
し、それぞれの撮像倍率での粒径測定範囲に違いを持た
せるとともに、各倍率の粒径測定範囲が互に部分的にオ
ーバーラップするようにしたことを特徴とする請求項１
記載の粒子画像分析装置。6. The image pickup means has means for selecting an image pickup magnification, and the particle size measurement ranges at the respective image pickup magnifications are made different from each other, and the particle size measurement ranges of the respective magnifications partially overlap each other. The method according to claim 1, wherein
The described particle image analyzer.
された粒子像から、それぞれ粒度頻度データを算出し、
それぞれの撮像倍率での試料分析量の違いにより粒度頻
度データを補正し、さらにそれぞれの撮像倍率での粒度
頻度データを加重平均法によって滑らかにつなぎ合わせ
ることを特徴とする請求項６記載の粒子画像分析装置。7. The calculation means calculates particle size frequency data from particle images captured at a plurality of types of imaging magnification,
7. The particle image according to claim 6, wherein the particle size frequency data is corrected according to the difference in the sample analysis amount at each imaging magnification, and the particle size frequency data at each imaging magnification is smoothly connected by the weighted average method. Analysis equipment.
像に対して単一の粒子であるか、複数個凝集した凝集粒
子であるかを識別する入力手段と、識別された単一の粒
子と凝集粒子の数の比を算出して表示手段に表示する手
段とをさらに備えたことを特徴とする請求項１記載の粒
子画像分析装置。8. The image analysis means includes an input means for identifying whether each particle image displayed collectively is a single particle or a plurality of aggregated particles, and a single identified single particle. The particle image analyzer according to claim 1, further comprising: a unit that calculates a ratio of the numbers of particles and agglomerated particles and displays the ratio on a display unit.
像に対して単一の粒子であるか、複数個凝集した凝集粒
子であるかを識別する入力手段をさらに備え、算出手段
は、単一の粒子として識別された粒子像だけを対象にし
て円相当径と円形度を算出することを特徴とする請求項
１記載の粒子画像分析装置。9. The image analysis means further comprises input means for identifying whether each particle image displayed collectively is a single particle or a plurality of agglomerated particles, and the calculation means comprises: The particle image analyzer according to claim 1, wherein the equivalent circle diameter and the circularity are calculated only for the particle image identified as a single particle.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP27145394A JP3411112B2 (en) | 1994-11-04 | 1994-11-04 | Particle image analyzer |
| US08/552,675 US5721433A (en) | 1994-11-04 | 1995-11-03 | Apparatus and method for analyzing particle images including measuring at a plurality of capturing magnifications |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP27145394A JP3411112B2 (en) | 1994-11-04 | 1994-11-04 | Particle image analyzer |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH08136439A JPH08136439A (en) | 1996-05-31 |
| JP3411112B2 true JP3411112B2 (en) | 2003-05-26 |
Family
ID=17500246
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP27145394A Expired - Lifetime JP3411112B2 (en) | 1994-11-04 | 1994-11-04 | Particle image analyzer |
Country Status (2)
| Country | Link |
|---|---|
| US (1) | US5721433A (en) |
| JP (1) | JP3411112B2 (en) |
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Also Published As
| Publication number | Publication date |
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| US5721433A (en) | 1998-02-24 |
| JPH08136439A (en) | 1996-05-31 |
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